Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: (A) Time-dependent changes in cytokine and chemokine production in LPS-stimulated THP-1 cells. After LPS (10 μg/ml) treatment for 15, 30, 60, or 120 min without any agents and with minocycline (50 μg/ml), tigecycline (50 μg/ml), or doxycycline (50 μg/ml), cytokines and chemokines were measured using Multi Plex according to the manufacturer’s protocols. * p <0.05 compared with LPS only at 60 min. ** p <0.05 compared with LPS only at 120 min. Mino: minocycline, Tige: tigecycline, Doxy: doxycycline. (B) The rate of cytokine and chemokine production in the THP-1 cell line compared to the production of cytokines and chemokines by LPS stimulation without tetracyclines. After LPS treatment (10 μg/ml) for 30, 60, 120 or 240 min without any agents and with minocycline (50 μg/ml), tigecycline (50 μg/ml), or doxycycline (50 mg/ml), cytokines and chemokines were measured with Multi Plex.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: Effects of minocycline, doxycycline, and tigecycline on the modulation of NF-κB, phospho-NF-κB, IKKα/β, phospho-IKKα/β, IκBα, and phospho-IκBα in LPS-stimulated THP-1 cells. THP-1 cells were incubated without or with 10 μg/ml LPS, or with LPS plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 30, 60, or 120 min. NF-κB, phospho-NF-κB, IKKα/β, phospho-IKKα/β, IκBα, and phospho-IκBα were assessed with Western blotting.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Incubation, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: Effects of tetracyclines (minocycline, doxycycline, and tigecycline) on the activation of phospho-ERK1/2 and phospho-p38 in LPS-stimulated THP-1 cells. THP-1 cells were incubated without or with 10 μg/ml LPS, or with LPS plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 30 or 60 min. Phospho-p38 and phospho-ERK1/2 were assessed with Western blotting.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Activation Assay, Incubation, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: SB203580, U0126 and BAY11-7082 suppressed TNF-α and IL-8 production in LPS-stimulated THP-1 cells on treatment with or without tetracyclines. THP-1 cells were pre-incubated by SB203580 (10 μM), U0126 (5 μM) and BAY11-7082 (5 μM) for 30 min, followed treatment without or with LPS (10 μg/ml), or with LPS (10 μg/ml) plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 60 min. TNF-α were measured with ELISA. * p <0.05, ** p <0.01, *** p <0.001 compared to the measurement without the inhibitor in the same group. Abbreviation; SB: SB203580, U: U0126, BAY: BAY11-7082.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: E3 ubiquitin ligase RNF128 promotes Lys63-linked polyubiquitination on SRB1 in macrophages and aggravates atherosclerosis

doi: 10.1038/s41467-025-57404-6

Figure Lengend Snippet: A Subset of cells from atherosclerotic aortas was annotated into nine clusters. B Location of cell clusters with Rnf128 expression on the t-SNE plot. C Cell clusters with the expression of Rnf128 ( Rnf128 + Mφ, left), high expression of Lyz2 ( Lyz2 hi Mφ, middle), and both of these two genes ( Rnf128 + Lyz2 hi Mφ, right). Mφ, macrophages. D The expression level of Rnf128 -expressing macrophage numbers during the process of atherosclerosis. E The number of macrophages with high expression of Rnf128 and Lyz2 in atherosclerotic aortas of mice fed WD for different durations. F Colocalization analysis via immunofluorescence of RNF128 and MOMA-2 (specific for monocytes and macrophages) expression in early and advanced atherosclerotic lesions of apolipoprotein E null (ApoE −/− ) mice fed a WD for 8 weeks and 20 weeks, respectively ( n = 8 per group, hereafter n = 8). Scale bar: 100 µm. G Western blotting images of RNF128 protein levels from whole aortas and quantitative analysis ( n = 6). H Colocalization analysis of RNF128 and MOMA-2 expression in early and advanced atherosclerotic lesions from coronary atheromatous plaques of humans ( n = 8). Scale bar: 100 µm. I Immunofluorescence of RNF128 in macrophages incubated with oxidized low-density lipoprotein (oxLDL, 75 µg/mL) for different time points ( n = 6). Scale bar: 20 µm. J Western blotting (left, n = 4) and quantitative PCR (right, n = 6) of RNF128 expression in macrophages treated with oxLDL for different time points. K Western blotting (left, n = 4) and quantitative PCR (right, n = 6) of RNF128 expression in macrophages treated with a concentration gradient of oxLDL for 24 h. L , M Western blotting (left, n = 4) and quantitative PCR analysis (right, n = 6) of RNF128 expression in RAW264.7 and THP-1-derived macrophages treated with time-dependent oxLDL, respectively. The “ n ” represents the number of biologically independent samples. Data were presented as mean ± SD, Shapiro–Wilk method tested that all data were normally distributed. Unpaired two-tailed Student’s t -test was used for ( G ). One-way ANOVA followed by the Dunnett post hoc test was used for the others. Adjusted P values were provided in case of multiple-group comparisons. Source data are provided as a Source Data file.

Article Snippet: The human monocyte leukemia cell line (THP-1, KeyGene BioTech, passage No. 5 to passage No. 10) was treated with phorbol 12-myristate 13-acetate (PMA, 100 nM; HY-18739, MCE, China) for 24 h for transformation into adherent macrophages.

Techniques: Expressing, Immunofluorescence, Western Blot, Incubation, Real-time Polymerase Chain Reaction, Concentration Assay, Derivative Assay, Two Tailed Test

Journal: Frontiers in Molecular Biosciences

Article Title: Nicotinamide Mononucleotide Alleviates LPS-Induced Inflammation and Oxidative Stress via Decreasing COX-2 Expression in Macrophages

doi: 10.3389/fmolb.2021.702107

Figure Lengend Snippet: NMN suppressed inflammatory cytokines in LPS-induced macrophages (A,C,E) The mRNA expressions of IL-6 and IL-1β were examined by RT-qPCR in untreated, LPS-treated, NMN-treated, LPS/NMN co-treated RAW264.7 cells, THP-1 cells and mouse peritoneal macrophages. (B,D,F) The ELISA results of IL-6 and IL-1β in untreated, LPS (100 ng/ml)-treated, NMN (500 µM)-treated and LPS-NMN-treated RAW264.7 cells, THP-1 cells and mouse peritoneal macrophages. The experiment was performed in triplicates and repeated three times. P values were calculated using one way ANOVA test. **** p < 0.0001, ** p < 0.01, * p < 0.05; mean ± SEM.

Article Snippet: For macrophage activation, THP-1 cells were treated with 10 ng/ml PMA (Beyotime, S1819) for 12 h to obtain PMA- activated THP-1 macrophages.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Nicotinamide Mononucleotide Alleviates LPS-Induced Inflammation and Oxidative Stress via Decreasing COX-2 Expression in Macrophages

doi: 10.3389/fmolb.2021.702107

Figure Lengend Snippet: NMN decreased cellular COX-2 expression in RAW264.7 cells, THP-1 cells and mouse peritoneal macrophages. (A) COX-2 levels from the proteomics data. (B,C) COX-2 expression in RAW264.7 cells was analyzed by western blotting. (D,E) COX-2 expression in THP-1 cells was analyzed by western blotting. (F,G) COX-2 expression in peritoneal macrophages was analyzed by western-blotting. The experiment was performed in triplicates and repeated three times. P values were calculated using one way ANOVA test. **** p < 0.0001, ** p < 0.01, * p < 0.05; mean ± SEM.

Article Snippet: For macrophage activation, THP-1 cells were treated with 10 ng/ml PMA (Beyotime, S1819) for 12 h to obtain PMA- activated THP-1 macrophages.

Techniques: Expressing, Western Blot